Uhn Lee

Neuroprotection of adenoviral-vector-mediated GDNF expression against kainic-acid-induced excitotoxicity in the rat hippocampus.

Glial-cell-line-derived neurotrophic factor (GDNF) is a potent survival factor for several types of neurons. In the present study, we examined the protective roles of adenoviral-vector-delivered GDNF (Ad-GDNF) in the hippocampus damaged by kainic-acid (KA)-induced excitotoxicity using GAD-67 immunoreactivity, immunoblot analysis, behavioral test, 5-bromo-2-deoxyuridine (BrdU) and TUNEL assay. Ad-GDNF was pre-inoculated into the KA-treated rat hippocampus 7 days before KA injection. Ad-GDNF resulted in the suppression of KA-induced tonic-clonic convulsions. In situ apoptosis assay demonstrated a significant reduction in apoptotic cells in the CA3 and dentate hilus regions of the Ad-GDNF-pre-inoculated rats (Ad-GDNF-KA), compared to the KA rats. Striking reductions in the density of GAD-67 neurons were also observed in the CA3 and dentate hilus regions of the KA rats. On the other hand, the number of GAD-67-positive cells was recovered to the control levels in the Ad-GDNF-KA rats. Immunoblot analysis further confirmed that GAD-67 and Bcl-2 expression increased in the Ad-GDNF-KA rats compared to KA rats. Taken together, these results suggest that Ad-GDNF may serve to control KA-induced hippocampal cell loss and behavioral seizure.

Tamoxifen-induced cell death and expression of neurotrophic factors in cultured C6 glioma cells.

The effects of ( Z)-2[ p -(1,2-diphenyl-1-butenyl)phenoxy]-N ,N -dimethylamine citrate (tamoxifen) on cell survival and the expression of neurotrophic factors (NTF) were investigated in rat C6 glioma cells (C6). C6 cells do not express the estrogen receptor. Cytotoxic effect was detected from 24 h after the treatment with 10 μM tamoxifen and increased with time in a dose-dependent manner. C6 cells treated with tamoxifen also displayed various morphological types such as elliptical, round and aggregated form. As the treatment time increased, the proliferation of C6 cells was reduced remarkably and most of them became the round or aggregated form.To examine the relationship of the expression of NTF and the cytotoxicity of tamoxifen, the mRNA level of brain-derived neurotrophic factor (BDNF), glial-derived neurotrophic factor (GDNF), and basic fibroblast growth factor (bFGF) was measured after 24 h treatment with tamoxifen by RT-PCR. The expression of mRNA of BDNF or GDNF in C6 cells treated with various concentrations of tamoxifen was comparable to controls. The expression of bFGF mRNA was significantly reduced in C6 cells treated with 10 or 15 μM tamoxifen. The results suggest that tamoxifen exerts cytotoxic effect on estrogen receptor-negative C6 cells through the inhibition of the transcription of bFGF.

Apoptosis and nestin expression in the cortex and cultured astrocytes following 6-OHDA administration

We used the dopaminergic neurotoxicant, 6-hydroxydopamine (6-OHDA), as a tool to characterize the origins of the astrocytic response to injury. Reactive astrocytes were examined by immunocyto- and histo-chemical visualization of nestin protein in the brain and cultivated cells. Following 6-OHDA (dose-dependent) treatment, the expression of nestin-like immunoreactive cells in the corpus callosum and cerebral cortex was increased compared with that of the control animals, indicating that a significant up-regulation of nestin protein occurred in these regions. In the corpus callosum and cerebral cortex, the majority of the nestin-like immunoreactive cells showed a distribution and pattern similar to those of the glial fibrillary acid protein (GFAP)-immunoreactive cells. Double immunofluorescence measurements showed that 100% of the nestin-like immunoreactive cells expressed GFAP-immunoreactive cells, indicating that these nestin-like immunoreactive cells belong to a reactive population of the astrocytes. In this study, we observed the morphological changes in the astrocytes following 6-OHDA administration, demonstrating that 6-OHDA induced injury leads to a rapid and transient up-regulation of nestin-like immunoreactivity in activated astrocytes.

Cytotoxicity of gold nanoparticles in human neural precursor cells and rat cerebral cortex.

Nanoparticles are promising tools for the advancement of drug delivery, medical imaging, and as diagnostic sensor. Medical nanodevices should develop miniaturization, because it would be injected into a human body. Gold nanoparticles (GNPs) with different sizes and shapes have therapeutic potential as a result of their small size, robust nature, excellent biocompatibility and optical properties. However, the application of GNPs as medical nanodevices it is necessary to know the biodegradation, biocompatibility, and development of surface coating which avoid the accumulation of nanoparticles. In this study, we carry out an in vitro toxicity and in vivo gene expression study using two kinds of GNPs. We found that GNPs toxicity is dependent on the dose or size administrated after the injected GNPs into the brain, and small particle size GNPs appeared more nestin expression compared to large particle size at short term implantation. These findings of toxicity of GNPs may play an important role in development of in vivo tools for the safety of GNPs.

Perspective in Nanoneural Electronic Implants With Wireless Power-Feed and Sensory Control

New medical device technology is essential for diagnosing, monitoring, and curing wide spectrum of diseases, anomalies, and inflictions. For neural applications, currently available devices are generally limited to either a curing or a probing function. In this paper, we review the technology requirements for a new neural probe and cure device technology currently under development. The concept of the probe-pin device that integrates the probes for neurochemistry, neuroelectricity, temperature, and pressure into a single embodiment with a wireless power transmission was designed for the purpose of deep brain feedback stimulation (DBFS) with in situ neural monitoring. The probe considered for monitoring neurochemistry is a microspectrometer. The feature and size of the microspectrometer are defined for the DBFS device. Two types of wireless power transmission technology were studied for the DBFS device operation. The test results of pig skin showed that both power transmission technologies demonstrated the feasibility of power feed through human tissue.

The change of the neuron-glia differentiation rate in human neural precursor cells (HPCs) and Ad-BDNF-/-GDNF-infected HPCs following the administration of a neurotoxin.

Neurotrophic factors promote the survival of various neurons, including peripheral autonomic and sensory neurons, as well as central motor and dopamine neurons, and it is expected that they could function as therapeutic agents for neurodegenerative disease. We examined the changes in the neuron-glia differentiation rate in normal human neural precursor cells (HPCs), Ad-BDNF- and Ad-GDNF-infected HPCs following their treatment with 6-OHDA. We isolated the precursor cells from the human fetal midbrain. To investigate the expression of differentiated cell markers within neurons and glia after 6-OHDA-induced toxicity in HPCs, immunocytochemistry was performed. Our results showed that the treatment with 6-OHDA (100, 200, 300, 400 and 500 microM) for 24 h decreased the viability of the HPCs in vitro. Among the growth factors tested, BDNF and GDNF protected the HPCs against 6-OHDA-induced toxicity. Approximately, 5.8+/-2.2% and 0.5+/-0.1% of the HPCs treated with 6-OHDA were positive for the neuron marker, MAP2, and the oligodendrocyte marker, GalC, respectively, while 13.8+/-3.2% and 1.1+/-0.36% of the Ad-BDNF- or Ad-GDNF-infected HPCs treated with 6-OHDA stained positive for MAP2 and GalC, respectively. These results suggest that cocktail therapy using human precursor cells (HPCs) and certain neurotrophic factors (BDNF, GDNF) provide direct protection against 6-OHDA-induced toxicity and has an effect on the differentiation rate.

Resorption of Autogenous Bone Graft in Cranioplasty: Resorption and Reintegration Failure

Objective Re-implantation of autologous skull bone has been known to be difficult because of its propensity for resorption. Moreover, the structural characteristics of the area of the defect cannot tolerate physiologic loading, which is an important factor for graft healing. This paper describes our experiences and results with cranioplasty following decompressive craniectomy using autologous bone flaps. Methods In an institutional review, the authors identified 18 patients (11 male and 7 female) in whom autologous cranioplasty was performed after decompressive craniectomy from January 2008 to December 2011. We examined the age, reasons for craniectomy, size of the skull defect, presence of bony resorption, and postoperative complications. Results Postoperative bone resorption occurred in eight cases (44.4%). Among them, two experienced symptomatic breakdown of the autologous bone graft that required a second operation to reconstruct the skull contour using porous polyethylene implant (Medpor®). The incidence of bone resorption was more common in the pediatric group and in those with large cranial defects (>120 cm2). No significant correlation was found with sex, reasons for craniectomy, and cryopreservation period. Conclusion The use of autologous bone flap for reconstruction of a skull defect after decompressive craniectomy is a quick and cost-effective method. But, the resorption rate was greater in children and in patients with large skull defects. As a result, we suggest compressive force of the tightened scalp, young age, large skull defect, the gap between bone flap and bone edge and heat sterilization of autologous bone as risk factors for bone resorption.

Nano-Scale Lateral Milling with Focused Ion Beam for Ultra-Smooth Optical Device Surface~!2009-12-16~!2010-02-18~!2010-04-21~!

The effect of the nano-scale lateral milling process using a focused ion beam (FIB) was studied in order to prepare a flat and smooth surface suitable for the growth of optical device structures such as a distributed Bragg reflector (DBR) mirror on a rough gallium nitride (GaN) surface. A high-quality, smooth, and flat surface is very essential for high precision space optics. It was fabricated using the nano-scale lateral milling process and was analyzed using an atomic force microscope. A regular geometric corrugation ripple with an amplitude of 18Å was achieved using a 30 KeV gallium ion-beam oriented at a normal angle of incidence operating at a beam current of 7nA. We suggest a simple engineering model for the formation of regular geometric ripples made by the serial cutting sequence of focused ion beam. This model can improve various device fabrications with a focused ion beam.

Mitochondrial DNA in patients with essential tremor

Essential tremor (ET) is one of the most common of the movement disorders. However, there has been little agreement in the neurological literature regarding diagnostic criteria for ET. It is not clear to what extent ET is associated with defects of mitochondrial DNA. In this study, we analyzed mitochondrial DNA (mtDNA) from the blood cells of the normal and ET patients using the long and accurate polymerase chain reaction (LA-PCR) and PCR. The large deletions were detected within several regions of mtDNA, but were not detected in the D-loop or CO I regions in ET patients. From our study, it is suggested that ET is a disorder showing a deficiency of mtDNA multicomplexes, and it also appears that mitochondrial dysfunction could be one of the causative factors of ET.

Miniaturized and Wireless Optical Neurotransmitter Sensor for Real-Time Monitoring of Dopamine in the Brain

Real-time monitoring of extracellular neurotransmitter concentration offers great benefits for diagnosis and treatment of neurological disorders and diseases. This paper presents the study design and results of a miniaturized and wireless optical neurotransmitter sensor (MWONS) for real-time monitoring of brain dopamine concentration. MWONS is based on fluorescent sensing principles and comprises a microspectrometer unit, a microcontroller for data acquisition, and a Bluetooth wireless network for real-time monitoring. MWONS has a custom-designed application software that controls the operation parameters for excitation light sources, data acquisition, and signal processing. MWONS successfully demonstrated a measurement capability with a limit of detection down to a 100 nanomole dopamine concentration, and high selectivity to ascorbic acid (90:1) and uric acid (36:1).